Functional maturation of human stem cell-derived neurons in long-term cultures

  • Differentiated neurons can be rapidly acquired, within days, by inducing stem cells to express neurogenic transcription factors. We developed a protocol to maintain long-term cultures of human neurons, called iNGNs, which are obtained by inducing Neurogenin-1 and Neurogenin-2 expression in induced pluripotent stem cells. We followed the functional development of iNGNs over months and they showed many hallmark properties for neuronal maturation, including robust electrical and synaptic activity. Using iNGNs expressing a variant of channelrhodopsin-2, called CatCh, we could control iNGN activity with blue light stimulation. In combination with optogenetic tools, iNGNs offer opportunities for studies that require precise spatial and temporal resolution. iNGNs developed spontaneous network activity, and these networks had excitatory glutamatergic synapses, which we characterized with single-cell synaptic recordings. AMPA glutamatergic receptor activity was especially dominant in postsynaptic recordings, whereas NMDA glutamatergic receptor activity was absent from postsynaptic recordings but present in extrasynaptic recordings. Our results on long-term cultures of iNGNs could help in future studies elucidating mechanisms of human synaptogenesis and neurotransmission, along with the ability to scale-up the size of the cultures.
Metadaten
Author:Rebecca S. Lam, Felix M. Töpfer, Phillip G. Wood, Volker BusskampORCiD, Ernst BambergGND
URN:urn:nbn:de:hebis:30:3-425858
DOI:https://doi.org/10.1371/journal.pone.0169506
ISSN:1932-6203
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/28052116
Parent Title (English):PLoS one
Publisher:PLoS
Place of publication:Lawrence, Kan.
Contributor(s):Han-Chiao Isaac Chen
Document Type:Article
Language:English
Date of Publication (online):2017/01/12
Date of first Publication:2017/01/04
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2017/01/12
Volume:12
Issue:(1): e0169506
Page Number:26
First Page:1
Last Page:26
Note:
Copyright: © 2017 Lam et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
HeBIS-PPN:415175305
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Angeschlossene und kooperierende Institutionen / MPI für Biophysik
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Sammlungen:Universitätspublikationen
Licence (German):License LogoCreative Commons - Namensnennung 4.0